CN112931207B - Seedling growing method of virus-free test-tube taro - Google Patents
Seedling growing method of virus-free test-tube taro Download PDFInfo
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- CN112931207B CN112931207B CN202110265912.1A CN202110265912A CN112931207B CN 112931207 B CN112931207 B CN 112931207B CN 202110265912 A CN202110265912 A CN 202110265912A CN 112931207 B CN112931207 B CN 112931207B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Abstract
The invention discloses a seedling method of virus-free test-tube taro, which comprises the following steps of S1: inducing detoxicated tube taro; s2: preparing a seedling culture substrate; the formula of the seedling culture substrate is grass peat: vermiculite to sand to soil =2:1, adding proper amount of tap water when mixing evenly; s3: subpackaging the prepared seedling substrate in a nutrition pot; the specification of the nutrition pot is 8 cm by 8 cm; 2/3 of the substrate is filled in each nutrition pot; s4: cleaning the detoxified test tube taro; on the day of setting the value, taking the detoxified taro out of the culture medium, cleaning the culture medium, removing roots and removing weak and irregular shapes; s5: planting and managing; and (2) performing fixed-value seedling raising of the test tube taro in the insect-proof gauze shed from ten days to ten days, watering after the field planting of the test tube taro is completed, thoroughly watering for the first time, watering for every 3 to 5 days later, and keeping the temperature in the shed at 15 to 20 ℃. The seedling method can ensure that the survival rate of the test-tube taro reaches 100 percent, and the plant grows robustly and has developed root system.
Description
Technical Field
The invention relates to the technical field of taro seedling culture, in particular to a seedling culture method of virus-free test-tube taro.
Background
Taro (Colocasia esculenta (L.) schott.) is perennial root herbaceous plant of taro (Colocasia) of Araceae, the fresh weight of the corm of taro contains 69.6-73.7% of starch, 1.75-2.3% of protein, 0.47-0.68% of lipid, 0.059-0.169% of calcium, 0.113-0.274% of phosphorus, 0.0042-0.0050% of iron, also contains 18 amino acids such as serine, aspartic acid, alanine and arginine, 19 trace elements such as K, zn and Mn, and vitamins B1, B2, D2, anthocyanin, sterol, catalase, carotene, water-soluble polysaccharide and heteropolysaccharide, etc., the corm is rich in starch, can be eaten fresh, and can be used for processing and producing starch and alcohol. The non-starch polysaccharide of taro can reduce the chance of suffering from rectal cancer and has the function of enhancing cellular immunity and humoral immunity to different degrees.
Taro is one of the ancient crops in China. Native China and tropical swamp zones of Indian Malaysia. The corm of the taro is rich in starch, is a main food source in many countries of the south pacific, and in feijia, tangca, cuk islands and other countries, the taro is not only a basic food, but also a stable channel for foreign exchange income. Taro has a long cultivation history in China and extremely wide distribution, wherein various types of taro mainly coexist in southern areas, and multi-seed taro mainly exists in Yangtze river watersheds. The taro can be eaten fresh and can be deeply processed. The processed products are exported to Japan, korea and southeast Asia countries. The total export amount of the taros all over the world in 2003 is 12.03 hundred million tons, the main export country is China, the annual export amount is 11.45 hundred million tons, and the taros account for 95.18% of the total export amount of the world. Taro is used as a main local product in China, and has wide prospects in further development of production and export industries.
Taro is generally infected by the taro mosaic virus after long-term vegetative propagation by utilizing corms. According to incomplete statistics, the planting area of the Chinese yam is about 200-300 ten thousand mu, and about 70 percent of the Chinese yam is infected by the mosaic virus of the yam. The yield of the taro infected with the virus is reduced by 30 to 70 percent. In addition, the conventional taro is large in seed consumption and low in propagation coefficient, and the seed consumption per mu is 100-150 kg, so that the popularization of new taro varieties is not facilitated. Severely restricts the production and export of the taro and the communication of international interspecific resources. The research of the detoxification test tube taro induction technology is the best way to solve the problem at present.
In the 70 s of the 20 th century, scientists in Japan, america, goss Dachaga, philippine and other countries successively studied the influence of the dasheen mosaic virus on the yield of the dasheen, and the detoxified seedling of the dasheen is produced by combining stem tip in vitro culture, seed rescue culture, stem tip in vitro culture and thermotherapy. The detoxicated test-tube plantlets are obtained successively from Cao Huanhuan of Shanghai and Bi Kehua of tobacco stage in the early 90 s of China. The test-tube plantlet transplanting survival rate is low, the cultivation technical requirement is high, and the method can not be directly applied to actual production. The invention of the detoxification test tube taro induction technology and the application thereof solve the problems existing in taro production and are mainly shown in the following steps: firstly, the virus-free test tube taro is obtained by tissue culture and rapid propagation of a taro stem tip growing point, thereby removing taro mosaic virus which seriously harms taro production, solving the problem of taro degeneration caused by virus disease, and improving the quality and yield of taro; secondly, the detoxified tube taro has small volume and high propagation speed and can be produced in an industrial way; thirdly, compared with the test-tube seedling of the taro, the test-tube taro is simple in seedling culture, high in survival rate and more storage-resistant, so that a more effective method is provided for the application of the taro tissue culture technology to the actual production; fourthly, the virus-free tube taro successfully eliminates diseases such as the dasheen mosaic virus and the like, and is the international and inter-regional taro germplasm resource communication.
The tube plantlet of taro needs to be unpackaged and hardened before seedling cultivation because the tube plantlet of taro is grown in an environment which is constant in temperature, constant in illumination, sterile and has complete nutrition supply and is greatly different from the external natural conditions, and the tube plantlet is removed and extremely easy to die without being fully hardened.
Generally, in practical application, the tube taro has advantages over tube seedlings, and as the tube taro is a dormant organ, the tube taro is convenient to transport, storage-resistant and small in path loss; the test-tube plantlet is easy to lose water and wither, so that the test-tube plantlet is inconvenient to transport at a long distance and the survival rate of the test-tube plantlet can be influenced after the test-tube plantlet is stored for a short time. Therefore, the virus-free tissue culture and rapid propagation of the taro are best to induce the taro into virus-free test tube and then carry out seedling culture.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a seedling method of detoxified test-tube taro, which can enable the survival rate of the test-tube taro to reach 100 percent through the selection of a substrate and a nutrition pot, and the test-tube taro has strong plant growth and developed root system.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a seedling raising method of virus-free test-tube taro is characterized by comprising the following steps,
s1: inducing detoxicated tube taro;
s2: preparing a seedling culture substrate;
s3: subpackaging the prepared seedling substrate in a nutrition pot;
s4: cleaning the detoxified test tube taro;
s5: and (5) planting and managing.
Further, the specific operation of the detoxified test tube taro induction in the step S1 comprises the following steps,
s101: obtaining a virus-free regenerated plant of the taro; inducing stem tips of the taros to generate cluster buds, selecting stem tip meristems cut to be 0.3-0.5 mm from the cluster buds, culturing, and inducing to form regenerated plants with 3-4 leaves and 4.5-5.0 cm high; detecting viruses such as dasheen leaves and the like on the regenerated plant, and reserving a negative material for removing the viruses for proliferation and rapid propagation to obtain a virus-free regenerated plant;
s102: placing the virus-free regenerated plant of the taro into a culture medium of 40ml per bottle for culturing for 10 days; the components of the culture medium comprise: MS + ZT 1.0 mg/L + G31.0 mg/L + NAA 0.3 mg/L + sucrose 3%/L + agar 6.0G/L;
s103: adding 30ml of 1/2-3/4 MS + 6-8% sucrose/L culture medium into the culture bottle, and continuously culturing for 15-20 days to obtain the detoxified test-tube taro with consistent uniformity.
Further, the sterilized seedling substrate is fully and uniformly mixed 15 days before the virus-free test tube taro is fixedly planted, and the mixture is covered and stacked by a plastic film.
Further, the formula of the seedling substrate is grass peat: vermiculite, sand and soil are =2:1, and a proper amount of tap water is added when the mixture is uniformly mixed.
Further, the specific operation of the step S3 includes that the nutrition pot can be filled after the seedling substrate is stacked for 15 days, and the specification of the nutrition pot is 8 cm by 8 cm; each of the nutrition bowls was filled with 2/3 of the medium, and was not filled and filled.
Furthermore, on the day of setting, the detoxified test tube taro is taken out of the culture bottle, the culture medium is cleaned, the root system is pulled out, the weak small and irregular shapes are removed, and the test tube taro is not damaged when the root system is pulled out.
Further, the concrete operation of the step S5 comprises the steps of carrying out test tube taro fixed-value seedling culture in an insect-proof gauze shed from late ten days of 2 months to early ten days of 3 months, poking a seedling culture substrate in a nutrition pot by using a bamboo stick to open a hole with the depth of 1-1.5 cm during fixed value, putting the test tube taro with top buds facing upwards into the nutrition pot, and slightly compacting the seedling culture substrate by hands;
watering after the field planting of the test tube taro, thoroughly watering for the first time, watering once every 3-5 days later, and keeping the temperature in the greenhouse at 15-20 ℃.
Furthermore, the test tube taro terminal buds for setting the value are full and complete and consistent in size, and the mass is not less than 0.6 g.
The beneficial effects of the invention are:
1. the virus-free test-tube taro seedling raising method does not need seedling hardening, and saves time. The virus-free test-tube plantlet needs to be acclimated for a week before seedling culture, because the virus-free test-tube plantlet grows in a culture room with constant temperature, constant illumination, sterility and complete nutrition supply, has great difference with the external natural environment condition, is removed without full acclimatization and is extremely easy to die. The virus-free test tube taro does not need to be hardened, is directly taken out from a constant-temperature, constant-illumination and sterile culture room before seedling culture, is cleaned and planted in a proper medium, does not influence the survival rate, saves the seedling hardening time for one week and reduces manpower and material resources.
2. The virus-free tube taro seedling method is simple in management, high in survival rate and cost-saving. The virus-free test tube plantlet needs to be covered with a film and a sunshade net for heat preservation, moisture preservation and sun protection in the early stage, the survival rate can reach 95 percent, and the virus-free test tube taro can reach 100 percent only by regular watering after field planting. The virus-free tube taro seedling raising management is simple to operate, high in survival rate, low in production cost and easy to popularize.
3. The virus-free test-tube taro seedling method disclosed by the invention has the advantages that the virus-free test-tube taro is storage-resistant and convenient to transport, can be popularized in a large area in taro production areas, and improves the market competitiveness of taro products. Because the detoxified test tube taro is a nutritional dormant organ, the method is proper and can be preserved for a long time without influencing the survival rate of the test tube taro; the test-tube plantlets are easy to lose water and wither no matter how to preserve the test-tube plantlets, the survival rate of the test-tube plantlets is reduced, and the test-tube plantlets can only be popularized and planted in accessory areas, so that the popularization area and efficiency are influenced. In addition, the detoxified test tube taro removes various viruses such as taro mosaic and the like, so that the quality and the yield of taro products are improved, the market competitiveness of the products is enhanced, and the economic benefit of unit area is improved.
4. The virus-free test-tube taro seedling raising method is convenient to transplant in a field and high in survival rate. When the virus-free test-tube taro seedlings are transplanted to a field with the length of 10-15 cm, the nutrition pots of 8*8 cm are torn and planted in the field with the matrix, the root systems of the taro seedlings are not damaged, the seedlings do not need to be revived, and the survival rate is high. If the plug seedling is used, the seedling needs to be pulled out, so that the root system of the seedling is damaged to a certain extent, the seedling needs to be revived after the transplanting, the survival rate is influenced, and the mixed matrix with the proportion is not easy to scatter during the transplanting, thereby being beneficial to the survival of the seedling.
5. The invention also discloses an induction method of the virus-free test-tube taro, in the induction method, the virus-free test-tube taro is obtained by tissue culture and rapid propagation of the stem tip growing point of the taro, thus successfully eliminating the taro mosaic virus seriously harming the production of the taro, solving the problem of taro seed sexual degradation caused by the virus disease of the taro and improving the quality and the yield of the taro; and the detoxified test tube taro is small in volume, light in weight and convenient to transport, and is suitable for the seedling culture method of the detoxified test tube taro in the application.
Drawings
FIG. 1 shows the growth of the virus-free regenerated plant of taro in MS + ZT 1.0 mg/L + G31.0 mg/L + NAA 0.3 mg/L + sucrose 3%/L + agar 6.0G/L medium for the first day of cultivation;
FIG. 2 shows the growth of the virus-free regenerated plant of taro in the invention on the 10 th day of cultivation in MS + ZT 1.0 mg/L + G31.0 mg/L + NAA 0.3 mg/L + sucrose 3%/L + agar 6.0G/L medium;
fig. 3 shows a growth condition of a virus-free regenerated plant of a taro on day 1 in a medium with an mg/L of MS + ZT 1.0 and G31.0 mg/L + NAA 0.3 mg/L + sucrose 3%/L + agar 6.0G/L medium with 1/2-3/4 MS + sucrose 6-8%/L;
FIG. 4 shows the growth of virus-free regenerated plants of the potatoes in the present invention, the MS + ZT 1.0 mg/L + G31.0 mg/L + NAA 0.3 mg/L + sucrose 3%/L + agar 6.0G/L medium is added with 1/2-3/4 MS + sucrose 6-8%/L medium on day 15;
FIG. 5 shows the result of comparing the growth of virus-free regenerated plants of dasheen in the same time without adding 1/2-3/4 MS + sucrose 6-8%/L medium, wherein MS + ZT 1.0 mg/L + G31.0 mg/L + NAA 0.3 mg/L + sucrose 3%/L + agar 6.0G/L medium is used for mg/L + sucrose 6-8%/L medium.
Detailed Description
In order to make those skilled in the art better understand the technical solution of the present invention, the following further describes the technical solution of the present invention with reference to the drawings and the embodiments.
The first embodiment is as follows:
in the embodiment, medium-sized detoxified test-tube taro of Orthosiphon Escharge No. 1 (the grading standards of different test-tube taros are that the weight of the large test-tube taro is more than or equal to 1g and less than 1g and the weight of the small test-tube taro is less than 0.5 g) is selected for seedling culture, and the specific seedling culture steps comprise:
s1: inducing detoxicated tube taro;
specifically, S101: obtaining a taro virus-free regeneration plant; inducing the stem tip of the taro to generate cluster buds, selecting stem tip meristem cut to be 0.3-0.5 mm from the cluster buds to culture, and inducing to form a regenerated plant with 3-4 leaves and 4.5-5.0 cm; detecting viruses such as dasheen leaves and the like on the regenerated plant, and reserving a negative material for removing the viruses for proliferation and rapid propagation to obtain a virus-free regenerated plant;
s102: placing the virus-free regenerated plant of the taro into a culture medium of 40ml per bottle for culturing for 10 days; the components of the culture medium comprise: MS + ZT 1.0 mg/L + G31.0 mg/L + NAA 0.3 mg/L + sucrose 3%/L + agar 6.0G/L; the growth of the virus-free regenerated plant in the culture medium at the 1 st day is shown in figure 1, and the growth of the virus-free regenerated plant at the 10 th day is shown in figure 2.
S103: adding 30ml of 1/2-3/4 MS + 6-8% sucrose/L culture medium into the culture bottle, and continuously culturing for 15-20 days to obtain the detoxified test-tube taro with consistent uniformity. The growth conditions of the virus-free regenerated plants of the taro at the moment of adding 1/2-3/4 MS + sucrose 6-8%/L and 15 days after adding into the culture bottle are respectively shown in the attached figures 3 and 4.
In the process of inducing the virus-free test tube taro, after culturing the virus-free regenerated plant for 10 days, adding a culture medium of 1/2-3/4 MS + sucrose 6-8%/L and a culture medium without 1/2-3/4 MS + sucrose 6-8%/L for continuous culture for the same time, comparing the growth conditions of the virus-free regenerated plant with that shown in the attached figure 5, wherein in the attached figure 5, (a) the culture medium of 1/2-3/4 MS + sucrose 6-8%/L is not added in a bottle, and (b) the culture medium of 1/2-3/4 MS + sucrose 6-8%/L is added in a bottle. As can be seen from the attached figure 5, the regenerated plants without the culture medium added with 1/2-3/4 MS + sucrose 6-8%/L have vigorous growth potential and grow slowly, while the regenerated plants added with the culture medium added with 1/2-3/4 MS + sucrose 6-8%/L gradually wither and grow rapidly. Generally, the regenerated plant without adding a culture medium of 1/2-3/4 MS and 6-8% sucrose/L can obtain the tube taro capable of growing seedlings within 40-50 days, and the regenerated plant with 1/2-3/4 MS and 6-8% sucrose/L can obtain the tube taro which is complete in shape, consistent in size and capable of growing seedlings within 25-30 days.
S2: preparing a seedling culture substrate;
specifically, 15 days before the virus-free test tube taro is fixedly planted, the disinfected seedling substrate is fully and uniformly mixed and covered and stacked by a plastic film; the formula of the seedling culture substrate is grass peat: vermiculite, sand and soil are =2:1, and a proper amount of tap water is added when the mixture is uniformly mixed.
S3: subpackaging the prepared substrate in a nutrition pot;
specifically, the seedling substrate can be stacked for 15 days to be filled into a nutrition pot, and the specification of the nutrition pot is 8 cm-8 cm; each of the nutrition bowls was filled with 2/3 of the medium, and was not filled and filled.
S4: cleaning the detoxified test tube taro;
specifically, on the day of setting the value, taking out the detoxified test tube taro from the culture medium, cleaning the culture medium, removing the root system, removing weak and small and irregular shapes, and not damaging the test tube taro when removing the root system.
S5: and (5) value setting and management.
In the last 3 months, performing constant-value seedling raising of the taro in the test tube in an insect-proof gauze shed, in the constant-value process, poking a seedling raising substrate in a nutrition pot through a hole with the depth of 1-1.5 cm by using a bamboo stick, putting the taro with a top bud upwards in the nutrition pot, and slightly compacting the seedling raising substrate by hands;
watering after the field planting of the test tube taro, thoroughly watering for the first time, watering once every 3-5 days later, and keeping the temperature in the greenhouse at 15-20 ℃.
Furthermore, the test tube taro terminal buds for setting the value are full and complete and consistent in size, and the mass is not less than 0.6 g.
Further, in this embodiment, a comparative test is performed on 5 different seedling raising substrates, where the 5 different seedling raising substrates are: (1) grass carbon: sand: soil =1:2:1; (2) grass carbon: sand: soil =1:1:1; (3) grass carbon: sand: soil =2:1:1; (4) grass carbon: perlite: soil =2:1:1; (5) grass carbon: perlite: sand: soil =2:1:1:1; 47 test tube taro plants are planted in each substrate. Planting in the middle and last ten days of 2 months, and investigating the survival rate, plant height, leaf length/width and rooting condition of the tube taro when transplanting in the last ten days of 4 months, wherein the results are shown in the following table 1.
TABLE 1 investigation results of different substrate seedling of tube taro
As can be seen from Table 1, the survival rate of the tube taro in the matrix (2) reaches 100%, the plant height is 4.7cm at most, and then the matrix (5) is added, the survival rate and the plant height are 92.9% and 4.3cm respectively, but the leaf length/width and the rooting condition of the tube taro in the matrix (5) are both larger and more than those of the matrix (2), namely the growth potential of the tube taro plant in the matrix (5) is better than that of the matrix (2), so that the matrix (5) is more suitable for seedling raising of the tube taro by integrating multiple factors.
Example two:
in the embodiment, medium-sized detoxified test-tube taro of Orthosiphon Escharge No. 1 (the grading standards of different test-tube taros are that the weight of the large test-tube taro is more than or equal to 1g and less than 1g and the weight of the small test-tube taro is less than 0.5 g) is selected for seedling culture, and the specific seedling culture steps comprise:
s1: inducing detoxicated tube taro;
the specific operation of this step is exactly the same as in the first embodiment.
S2: preparing a seedling culture substrate;
specifically, 15 days before the virus-free test tube taro is fixedly planted, the disinfected seedling substrate is fully and uniformly mixed and covered and stacked by a plastic film; the formula of the seedling raising substrate is grass peat: vermiculite to sand to soil =2:1, adding proper amount of tap water when mixing evenly.
S3: subpackaging the prepared substrate in a nutrition pot;
specifically, the seedling substrate can be stacked for 15 days to be filled into a nutrition pot, and the specification of the nutrition pot is 8 cm by 8 cm; each of the nutrition bowls was filled with 2/3 of the medium, and was not filled and filled.
S4: cleaning the detoxified test tube taro;
specifically, on the day of setting the value, taking out the detoxified test tube taro from the culture medium, cleaning the culture medium, removing the root system, removing weak and small and irregular shapes, and not damaging the test tube taro when removing the root system.
S5: and (5) value setting and management.
In the last 3 months, performing constant-value seedling raising of the taro in the test tube in an insect-proof gauze shed, in the constant-value process, poking a seedling raising substrate in a nutrition pot through a hole with the depth of 1-1.5 cm by using a bamboo stick, putting the taro with a top bud upwards in the nutrition pot, and slightly compacting the seedling raising substrate by hands;
watering after the field planting of the test tube taro, thoroughly watering for the first time, watering once every 3-5 days later, and keeping the temperature in the greenhouse at 15-20 ℃.
Furthermore, the test tube taro terminal buds for setting the value are full and complete and consistent in size, and the mass is not less than 0.6 g.
Further, in this embodiment, a comparative test is performed on 6 different seedling raising containers, and the different seedling raising containers in 6 are respectively: and (3) plug disc: (1) 54 holes (2) and (70 holes (3); a nutrition pot: (4) 8*8 cm (5, 9 cm (6, 10, 8 cm; the plug tray is processed 3 times every time and is repeated 3 times; the nutrition pots were treated 25 times and repeated 3 times. Planting in the middle and last ten days of month 2, and examining the survival rate of the tube taro when transplanting in the last ten days of month 4, wherein the results are shown in Table 2.
TABLE 2 investigation results of different seedling raising containers for tube taro
As can be seen from Table 2, the survival rate of the test tube taro in the nutrition pot is higher than that in the plug tray, and the survival rate in the No. 4 nutrition pot reaches 100 percent, and then the No. 5 nutrition pot. (4) (5) plants in the nutrition pots grow strongly, the root systems are developed, the plants are easier to transplant into a field, the nutrition pots are only required to be torn and fixedly planted together with the substrate, seedlings are not damaged, the roots are not damaged, the plants are easy to survive, and the operation is convenient; when the plants in the plug tray are transplanted, the seedlings need to be pulled out, and the process can damage the seedlings and root systems and influence the survival of the seedlings; in addition, the nutrition body (4) has smaller volume than the nutrition bowls (5) and (6), and the substrate is saved, so that the nutrition bowl of 8*8 cm for seedling culture of the tube taro is better.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (3)
1. A seedling raising method of virus-free test-tube taro is characterized by comprising the following steps,
s1: inducing detoxicated tube taro;
s2: preparing a seedling culture substrate;
s3: subpackaging the prepared seedling substrate in a nutrition pot;
s4: cleaning the detoxified test tube taro;
s5: planting and managing;
the specific operation of the detoxified test tube taro induction in the step S1 comprises the following steps,
s101: obtaining a taro virus-free regeneration plant; inducing the stem tip of the taro to generate cluster buds, selecting stem tip meristem cut to be 0.3-0.5 mm from the cluster buds to culture, and inducing to form a regenerated plant with 3-4 leaves and 4.5-5.0 cm; detecting viruses such as dasheen leaves and the like on the regenerated plant, and reserving a negative material for removing the viruses for proliferation and rapid propagation to obtain a virus-free regenerated plant;
s102: placing the virus-free regenerated plant of the taro into a culture medium of 40ml per bottle for culturing for 10 days; the components of the culture medium comprise: MS + ZT 1.0 mg/L + G31.0 mg/L + NAA 0.3 mg/L + sucrose 3%/L + agar 6.0G/L;
s103: adding 30ml of 1/2-3/4 MS + 6-8% sucrose/L culture medium into the culture bottle, and continuously culturing for 15-20 days to obtain detoxified test-tube taro with consistent uniformity;
the specific operation of the step S2 comprises the following steps of fully mixing the sterilized seedling substrate 15 days before the field planting of the virus-free colocasia esculenta, covering and stacking the seedling substrate by using a plastic film, wherein the formula of the seedling substrate is turf: perlite: sand: soil =2:1, adding proper amount of tap water when evenly mixing;
the specific operation of the step S3 comprises the steps that the nutrition pot can be filled after the seedling substrate is stacked for 15 days, and the specification of the nutrition pot is 8 cm by 8 cm; 2/3 of the substrate is filled into each nutrition pot, and the nutrition pots cannot be filled or are not filled;
the concrete operation of the step S5 comprises that in the last ten days of 2 months to the last 3 months, test tube taro fixed-value seedling raising is carried out in an insect-proof gauze shed, when the value is fixed, a bamboo stick is used for poking a seedling raising substrate in a nutrition pot open a hole with the depth of 1 cm-1.5 cm, the test tube taro with apical bud upwards is placed in the nutrition pot, and the seedling raising substrate is lightly compacted by hands;
watering after the field planting of the test tube taros, wherein the water is thoroughly watered for the first time, and then is watered every 3 to 5 days, and the temperature in the greenhouse is kept between 15 and 20 ℃.
2. The method for growing seedlings of virus-free colocasia esculenta as claimed in claim 1, wherein: the specific operation of the step S4 comprises the following steps of taking out the detoxified test tube taro from the culture bottle on the day of setting, cleaning the culture medium, removing the root system, removing weak and small and irregular shapes, and not damaging the test tube taro when removing the root system.
3. The method for growing seedlings of virus-free colocasia esculenta as claimed in claim 1, wherein: the test tube taro terminal buds for setting the value are full and complete and consistent in size, and the mass is not less than 0.6 g.
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